success rate of cervical cancer treatment is a big challenge in the me - PDF document

1 3504 success rate of cervical cancer tre
3504 success rate of cervical cancer treatment is a big challenge in the medical �eld. targeted therapy is the �rst choice in clinic8,9. the curative effect of current molecular targeting anti-apoptotic proteins, such as survivin and apollon on cervical cancer, is still not satisfactory. therefore, it is urgently required to explore more effective molecular targets for the treatment of cervical cancer in clinic. Paclitaxel is a kind of important �rst-line anticancer drug for the treatment of cervical cancer Abstract.OBJECTIVE: Paclitaxel is one of the common anticancer drugs in the treatment European Review for Medical and Pharmacological Sciences Department of Obstetrics, Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, ChinaDepartment of Pediatrics, Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, ChinaDepartment of Gynecology and Obstetrics, Huangdao Qingdao District Wang Tai Center Hospital, Qingdao, ChinaDepartment of Gynecology, Qingdao Haici Hospital, Qingdao, Shandong Province, ChinaDown-regulation of survivin enhances paclitaxel-induced Hela cell apoptosis 3505 that survivin may play an important role in Hela cells apoptosis. However, the exact mechanism by how survivin induces Hela cells apoptosis remains unclear. the aim of the present study was to investigate the speci�c regulatory mechanism of survivin on Hela cells apoptosis.Cell apoptosis detection kit was purchased from Beyotime (Shanghai, China). Mtt, lipofectamine reagent escort, and rabbit anti-human survivin primary antibody were purchased from Sigma-aldrich (St. Louis, MO, USa). Survivin sirna and control sirna were designed and synthetized by Genepharma (Shanghai, China). the sequences were 5’GCGatGattGGttatCGt3’, and 5’aaaataGGtCatCatGGt3’, respectively. Dulbecco’s Modi�ed eagle Medium (DMeM) was provided by Beijing Dingguo C

2 hangsheng Biotechnology Co. Ltd (Beijing
hangsheng Biotechnology Co. Ltd (Beijing, China).Hela cells were purchased from atCC. the cells were resuscitated and routinely cultured. Paclitaxel at 2 μmol/L or dimethyl sulfoxide (DMSO) was applied to treat the cells for 48 h.the cells were seeded and transfected using survivin sirna and Lipofectamine reagent escort according to the manual.Cell membrane potential speci�c dye tMre and cell membrane phosphatidylserine speci�c dye FItC-annexin V were applied to test the apoptosis of cervical cancer cell line by �ow cytometry. the cells were seeded into 96-well plate at a density of 10.000 cells/well for analysis of cell proliferation by methylthiazolyl tetrazolium bromide (Mtt) assay.the cells were cracked on ice for 10 min and added into the 96-well plate. then, the chromophoric substrate was added to the plate for measuring the caspase-3 activity.the cells were treated with lysis buffer and separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PaGe). Primary antibodies for survivin and actin were applied for detection.all data analysis was performed with SPSS18.0 software (SPSS Inc., Chicago, IL, USa). Data were represented as mean ± standard deviation (SD). Student’s -test was performed for comparing the difference between two groups. One-way anOVa and Dunnet post hoc test were performed to assess the statistical signi�cance among multiple treatment groups. a signi�cant difference was depicted as < 0.05.the impacts of paclitaxel on Hela cell survival and proliferation were tested by Mtt assay. as shown in Figure 1, paclitaxel signi�cantly suppressed Hela cell growth ( < 0.05), suggesting it affects the proliferation of Hela cells. Mitochondrial membrane potential of Hela cells was evaluated by �ow cytometry after paclitaxel treatment. It was found that paclitaxel obviously declined mitochondrial memb

3 rane potential of Hela cells ( < 0.05) (
rane potential of Hela cells ( < 0.05) (Figure 2), indicating that paclitaxel may induce the apoptosis of Hela cells.to con�rm whether paclitaxel induces apoptosis of Hela cells, we measured the phosphati Figure 1. Paclitaxel inhibited Hela cells proliferation. * < 0.05 vs. control. 3506 dylserine exposure of Hela cells after paclitaxel treatment. results showed that after paclitaxel intervention, Hela cells exhibited markedly increased phosphatidylserine exposure ( < 0.05) (Figure 3), revealing that paclitaxel induces Hela cell apoptosis. Considering paclitaxel-induced apoptosis of Hela cells, caspase-3 activity was also measured. as shown in Figure 4, caspase-3 activity in Hela cells was obviously enhanced after paclitaxel treatment. Western blot was adopted to assess survivin protein expression in Hela cells. It was revealed that survivin protein level was down-regulated in Hela cells after treated with paclitaxel (Figure 5).as shown in Figure 6, survivin level in Hela cells was suppressed after survivin sirna transfection, whereas, caspase-3 activity was enhanced in survivin sirna transfected Hela cells treated with paclitaxel, revealing that down-regulation of survivin facilitated paclitaxel-induced Hela cells apoptosis.Paclitaxel is an important anti-cancer drug in clinic. In this study, we discussed the impacts of paclitaxel on the growth and apoptosis of cervical cancer cell line Hela. Firstly, we explored the influence of paclitaxel on cervical cancer Hela cells growth and survival and showed paclitaxel significantly decreased the proliferation of Hela cells, which is consistent with previous study. to evaluate the effect of Figure 2. Paclitaxel reduced cell membrane potential in Hela cells. < 0.05 vs. control. F. Gu, L. Li, Q.-F. Yuan, C. Li, Z.-H. Li 3507 paclitaxel on Hela cells apoptosis, flow cytometry and caspase-3 activity detection revealed that Hela cells exhibited mitochondr

4 ial membrane potential reduction and pho
ial membrane potential reduction and phosphatidylserine exposure, suggesting that paclitaxel-induced Hela cells apoptosis possibly through activation of caspase-3, which was in accordance with previous studies17,18. However, va Figure 3. Paclitaxel-induced Hela cell phosphatidylserine exposure. * < 0.05 vs. control. Figure 4. Paclitaxel activated caspase-3 in Hela cells. *0.05 vs. control. Figure 5. Paclitaxel downregulated survivin protein expression in Hela cells. 3508 rious apoptosis indexes and detection methods showed that under the similar concentrations of paclitaxel (2 μmol/L), Hela cells presented a higher sensitivity compared with other cancer cells. It may be caused by the fact that different cells have the diverse sensitivity to the same chemical drug. apoptosis pathway mainly activates caspase-8, while mitochondria-mediated intrinsic apoptosis pathway may activate caspase-3/7. In this study, paclitaxel activated caspase-3 of Hela cells, suggesting that the apoptosis of Hela cells induced by paclitaxel was possibly through mitochondria-mediated intrinsic apoptosis pathway, which was in agreement of previous investigations20,21. as an inhibitor of apoptosis protein, survivin plays a critical role in apoptosis, necroptosis, autophagy and nF-κB signaling pathway. Our results revealed that paclitaxel treatment significantly increased the apoptotic rate of Hela cells with down-regulation of survivin, indicating that knockdown of survivin contributes to cancer therapy. Based on our data, several investigations could be carried out in the future. Clinical cervical cancer specimens in different stages could be collected to detect survivin expression to evaluate whether survivin is associated with clinical stages. Different types of paclitaxel and similar drugs could be adopted to clarify the specific target and mechanism of paclitaxel on cervical cancer, thus providing more valuable information for

5 clinical practice. However, future studi
clinical practice. However, future studies are required to confirm our findings using survivin knockout mice. Paclitaxel-treated Hela cells display reduced proliferation and increased apoptosis. Moreover, down-regulation of survivin facilitates paclitaxel-induced apoptosis of cervical cancer cell line Hela cells.Conflict of interestthe authors declare no con�icts of interest.1) RANCO EL, SCHLECHTF, SSLOW D . The epidemiology of cervical cancer. Cancer J 2003; 9: 348-359. ONGJ 3 Management of metastatic cervical cancer: review of the literature. J Clin Oncol 2007; 25: 2966-2974. BEANU O Molecular pathogenesis of cervical cancer. Cancer Biol Ther 2011; 11: 295-306. ANG J, ZHANG, J S. Epigenetics and cervical cancer: from pathogenesis to therapy. Tumour Biol 2014; 35: 5083-5093. SJ, PS, JENSO Cervical cancer: etiology, pathogenesis, treatment, and future vaccines. Asian Pac J Cancer Prev 2002; 3: 207-214. OOREEE, WARK JD, OPPER JL, RBA, ARAN S The roles of genetic and environmental factors on risk of cervical cancer: a review of classical twin studies. Twin Res Hum Genet 2012; 15: 79-86.7) ERATA-ZARAGOZ O, ERMURA V, P - RE-PENC, SAR-L J, MEERIDARNA V. Targeted treatments for cervical cancer: a review. Onco Targets Ther 2012; 5: 315-328. AGUR F, SERGENTANISTNHRYSIOS D, FILIPIARTCH Molecularly targeted therapies in cervical cancer. A systematic review. Gynecol Oncol 2012; 126: 291-303. ENNAT, FOLI, ZAR Targeting survivin in cancer therapy. Expert Opin Ther Targets 2008; 12: 463-476.10) RTEENNEEBLI L, FUNG Systemic therapy for recurrent, persistent, or metastatic cervical cancer: a clinical practice guideline. Curr Oncol 2015; 22: 211-219.11) RZIJ, HORSE F, EESE, VED, BJERKIG, DAHRNOLDUNDESE . Proliferation, migration and invasion of human glioma cells exposed to paclitaxel (Taxol) in vitro. Br J Cancer 1997; 75: 1744-1752.12) IRUPPATHI, SONG BRGFELD M, SSSIK Gp60 activation mediates albumin t

6 ranscytosis in endothelial cells by tyro
ranscytosis in endothelial cells by tyrosine kinase-dependent pathway. J Biol Chem 1997; 272: 25968-25975.Figure 6. Paclitaxel-induced Hela cells apoptosis through activating survivin. < 0.05 vs. control. ** < 0.01 vs. control. F. Gu, L. Li, Q.-F. Yuan, C. Li, Z.-H. Li 3509 13) HIGEMAT, OYA, MOTO, S, HARA M, ITANI, KA M . the ef�cacy and safety of preoperative chemotherapy with triweekly abraxane and cyclophosphamide followed by 5-�uorouracil, epirubicin, and cyclophosphamide therapy for resectable breast cancer: a multicenter clinical trial. Clin Breast Cancer 2015; 15: 110-116.14) AI M.S. Food and drug administration approves paclitaxel protein-bound particles (Abraxa) in combination with gemcitabine as �rst-line treatment of patients with metastatic pancreatic cancer. JOP 2013; 14: 686-688.15) EE M, MANIKHAM, S, ANAEV B, AKHONM, BHARAWKIN MJ. Abraxane, a novel cremophor-free, albumin-bound particle form of paclitaxel for the treatment of advanced non-small-cell lung cancer. Ann Oncol 2006; 17: 1263-1268.16) TIRI D Survivin, versatile modulation of cell division and apoptosis in cancer. Oncogene 2003; 22: 8581-8589.17) ANGTHANGS, SOONGYK . Paclitaxel-induced cell death: where the cell cycle and apoptosis come together. Cancer 2000; 88: 2619-2628.18) ONEFE, EDE, ESSMANN F, SCHUTHOFF, DORK B, DANIELPT Paclitaxel-induced apoptosis in BJAB cells proceeds via a death receptor-independent, caspases-3/-8-driven mitochondrial ampli�cation loop. Oncogene 2003; 22: 2236-2247.19) traSSer, O’Cnn L, ⁄i硩t⁖M.⁁poptos楳⁳ignaling⸠ann甠re瘠Bioche洠2000㬠69㨠217-24㔮2〩 UTUK, LTAI . Alteration of the mitochondrial apoptotic pathway is key to acquired paclitaxel resistance and can be reversed by ABT-737. Cancer Res 2008; 68: 7985-7994.21) ONLE DJ, D D . Apoptosis and taxol therapy. Cancer Biol Ther 2002; 1: 118-120. Survivin mediates paclitaxel effect in